PROJECT SUMMARY/ABSTRACT This proposal requests funds for a user-friendly cell sorter to be incorporated into the Research Flow Cytometry Core (RFCC) that serves investigators at Cincinnati Children's Hospital Medical Center (CCHMC) and University of Cincinnati College of Medicine (UCCOM). The Sony SH800 cell sorter has been selected as it best fulfills the needs of our NIH funded users. A dramatic expansion of research in the past 10 years at CCHMC and UCCOM, coupled with an increase in novel technologies downstream of cell sorting, such as single cell gene expression and CRISPR, have increased the number of investigators utilizing single cell analysis. This is reflected in an increase of investigators using RFCC cell sorting services from 58 investigators in 2011, to 107 in 2015. Current RFCC cell sorters are being utilized at greater than 80% capacity, limiting accessibility. In addition, our major users are in need of access to this technology 24/7 as their translational projects involve patient samples, for which time of attainability is not easily predicted. The current cell sorters are complex instruments that require highly trained cell-sorting operators. These factors indicate an increased need for (1) a user-friendly instrument for sorts that need to be performed by users outside of normal operating hours, and (2) unassisted sorting for either simple fluorescent protein or 2-6 color immune-phenotyping panels which would alleviate the pressure on the current instrumentation in our facility. The instrument will support and enhance existing NIH-funded projects and will foster avenues of research that will likely lead to future funded projects. In this application, 18 investigators (7 major and 11 minor users, with 17 users funded by their own NIH grants) are identified whose projects require increased accessibility to cell sorting. The major users projects span a broad spectrum of biologic queries requiring the use of cell sorting, including: (1) immunologic regulation of biliary atresia; (2) immunobiology of Infection with intracellular pathogens; (3) fundamental mechanisms of hematopoietic stem cell self-renewal; (4) transcriptional control of hematopoiesis and cancer; (5) homeostasis and function of T cells; (6) macrophage based gene therapy for hereditary Pulmonary Alveolar Proteinosis; (7) natural killer cell contributions to antiviral immunity and vaccine efficacy. The incorporation of the SH800 into our current cell sorter repertoire will complement and enhance the operation of the RFCC and maximally benefit investigators in this application, as well as the entire user base in the CCHMC and UCCOM research community.